Fibronectin (Fn) is a 55OkDa glycoprotein found in most body fluids and in fibrillar form in most tissues where it mediates adhesion, spreading and migration of many cell types. It plays a major role in embryogenesis, cancer, wound healing, tissue remodeling, hematopoiesis, blood coagulation and fibrinolysis. Fn also facilitates removal of damaged tissue and clot remnants from circulation by phagocytes. Many bacteria and parasites are thought to use Fn as a means of colonizing tissue. Fn is a """"""""mosaic"""""""" protein composed of homologous modules (domains) of three different types that are also found in other proteins. The long range goal of this research is to understand the structure and function of each module in Fn, to know which ones are involved in the multiplicity of interactions and which amino acids within those modules participate directly in the recognition process. This will be accomplished by isolating ever smaller fragments by classical or recombinant techniques, characterizing their interactions and folding properties by calorimetry and fluorescence, and determining as much as possible about their structure.This application emphasizes the self- interactions that govern the supertertiary structure of Fn in solution and its transformation into an insoluble fibrillar component of the extracellular matrix. In addition we wish to continue our characterization of hetero-interactions with selected macromolecules (collagen, fibrinogen, heparin, S.aureus peptides) and to collaborate with others to elucidate the 3D structure of various fragments. The knowledge gained will provide a better understanding of the many functions of Fn and may suggest improved methods of intervention in disease processes.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL021791-22
Application #
2901030
Study Section
Molecular and Cellular Biophysics Study Section (BBCA)
Project Start
1978-01-01
Project End
2001-03-31
Budget Start
1999-04-01
Budget End
2000-03-31
Support Year
22
Fiscal Year
1999
Total Cost
Indirect Cost
Name
American National Red Cross
Department
Type
DUNS #
003255213
City
Washington
State
DC
Country
United States
Zip Code
20006
Ingham, Kenneth C; Brew, Shelesa A; Erickson, Harold P (2004) Localization of a cryptic binding site for tenascin on fibronectin. J Biol Chem 279:28132-5
Ingham, K C; Brew, S A; Migliorini, M (2002) Type I collagen contains at least 14 cryptic fibronectin binding sites of similar affinity. Arch Biochem Biophys 407:217-23
Katagiri, Yasuhiro; Ingham, Kenneth C (2002) Enhanced production of green fluorescent fusion proteins in a baculovirus expression system by addition of secretion signal. Biotechniques 33:24-6
Makogonenko, Evgeny; Tsurupa, Galina; Ingham, Kenneth et al. (2002) Interaction of fibrin(ogen) with fibronectin: further characterization and localization of the fibronectin-binding site. Biochemistry 41:7907-13
Akimov, S S; Krylov, D; Fleischman, L F et al. (2000) Tissue transglutaminase is an integrin-binding adhesion coreceptor for fibronectin. J Cell Biol 148:825-38
Yakovlev, S; Makogonenko, E; Kurochkina, N et al. (2000) Conversion of fibrinogen to fibrin: mechanism of exposure of tPA- and plasminogen-binding sites. Biochemistry 39:15730-41
Bloom, L; Ingham, K C; Hynes, R O (1999) Fibronectin regulates assembly of actin filaments and focal contacts in cultured cells via the heparin-binding site in repeat III13. Mol Biol Cell 10:1521-36
Litvinovich, S V; Brew, S A; Aota, S et al. (1998) Formation of amyloid-like fibrils by self-association of a partially unfolded fibronectin type III module. J Mol Biol 280:245-58
Medved, L; Litvinovich, S; Ugarova, T et al. (1997) Domain structure and functional activity of the recombinant human fibrinogen gamma-module (gamma148-411). Biochemistry 36:4685-93
Ingham, K C; Brew, S A; Huff, S et al. (1997) Cryptic self-association sites in type III modules of fibronectin. J Biol Chem 272:1718-24

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